Advantages of Wild Diploid Solanum Species Over Cultivated Diploid Relatives in Potato Breeding Programs

For breeding programs of the tetraploid potato (Solanum tuberosum), both wild and cultivated diploid relatives are valuable sources of genetic diversity. While both types of germplasm are used in breeding programs, there are several advantages to using wild relatives. Diploid relatives are typically crossed with haploids (2n = 2x = 24) from tetraploid S. tuberosum to improve daylength adaptation. Most haploids are male sterile, so they are typically used as female parents. Cultivated diploids, such as members of the Phureja Group, produce male sterile hybrids when crossed as females to haploids; wild relatives, such as S. tarijense, often produce male fertile hybrids. Tuber yield following crosses of haploids to cultivated or wild relatives is often high. However, cultivated relatives generally produce hybrids with a high set of small tubers; hybrids from wild relatives are variable, but many are similar to cultivars in tuber size and set. While tubers of hybrids from cultivated relatives are typically rough, with deep eyes and raised internodes, those from wild relatives are often smooth. Tuber dormancy in hybrids with cultivated relatives is generally short, while that in hybrids with wild species is longer, allowing for storage over winter. Finally, resistance to several major diseases and stresses has been found in wild species and their hybrids with S. tuberosum haploids. The desirable traits in hybrids are transmitted to tetraploids via unilateral sexual polyploidization (4x × 2x or 2x × 4x crosses in which the diploid parent produces 2n gametes). Wild Solanum species are recommended for use in potato breeding programs as sources of genetic diversity that can be adapted easily following hybridization with S. tuberosum haploids.     

The introgression of 2× 1EBN <Emphasis Type="Italic">Solanum</Emphasis> species into the cultivated potato using <Emphasis Type="Italic">Solanum verrucosum</Emphasis> as a bridge

Wild diploid Solanum species with an endosperm balance number (EBN) of one are a rich source of disease resistance, pest resistance, and tuber quality traits. They are not sexually compatible with 4× 4EBN or 2× 2EBN forms of the cultivated potato. In this study, the wild 2× 2EBN Mexican species S. verrucosum (ver) was used as a bridge species to access 2× 1EBN germplasm. Hybrids (V1) were created between ver as a female parent and the 1EBN species S. cardiophyllum, S. chancayense, S. commersonii, and S. trifidum. Most of the V1 hybrids were successfully crossed as females to cultivated 2× 2EBN clones. However, due to stylar barriers, reciprocal crosses were unsuccessful. Additional studies are underway to determine the effectiveness of ver as a bridge to introgress specific traits from 2× 1EBN germplasm into the cultivated potato.     

Cytological mechanisms of 2n pollen formation in the wild potato Solanum okadae and pollen-pistil relations with the cultivated potato, Solanum tuberosum

Solanum okadae Hawkes et Hjert is a wild diploid potato species endemic to Argentina and Bolivia, of potential breeding value. However, no genetic studies have been carried out with this species and its crossability relations with the common tetraploid potato, S. tuberosum L. ssp. tuberosum, are unknown. Furthermore, accessions from both countries differed in their morphological phenotypes. To ascertain the feasibility of incorporating this wild germplasm into cultivated potato, 2n pollen screening was carried out in 10 accessions and families derived from crosses between accessions; also, pollen-pistil compatibility relations were studied in reciprocal interspecific crosses. Plants of four of the 10 accessions produced 2n pollen (0.1 to 5.0%) and 4n pollen (0.0 to 3.0%). Parallel and tripolar spindles at Anaphase II were the cytological mechanisms involved in 2n pollen formation; lack of chromosome migration in both meiotic divisions originated the 4n pollen. Both full compatibility and incompatibility at various sites along the pistil were observed in the S. tuberosum × S. okadae combinations; most reciprocal combinations were incompatible. Compatible genotypes produced 2n pollen. However, only a few seeds were obtained and chromosome counts could not be carried out in the hybrid progeny because seedlings died at an early stage. The pollen-pistil barriers are incomplete and can be circumvented by the appropiate choice of parents. The identification of the post-zygotic barriers will be the focus of further studies.     

Glycoalkaloid content and chemical composition of potatoes improved with nonconventional breeding approaches

This paper reports the results of chemical analyses performed on two distinct groups of new potato genotypes. The first group contained five clones transformed with the gene ech42 encoding for an endochitinase. The second included 21 interspecific hybrids between the cultivated potato Solanum tuberosum and the wild species S. commersonii, obtained either by somatic fusion or by sexual hybridization. Tubers from transgenic plants were analyzed for several morphological and biochemical parameters to ascertain the substantial equivalence between the transgenic genotypes and the original cultivar Désirée. The interspecific hybrids were analyzed for the same parameters in order to identify genotypes with novel improved chemical characteristics and with low levels of glycoalkaloids deriving from the wild species and potentially hazardous to human health. For transgenic tubers, the results provided evidence that indicates the substantial equivalence between the transgenic genotypes and the cultivated control for the considered traits. The results suggest that chitinase gene insertion did not alter other metabolic pathways of potato tubers and did not cause unintentional pleiotropic effects. As far as interspecific hybrids are concerned, wide variability for all of the parameters analyzed was found. For some useful traits (e.g., soluble solids and proteins, dry matter content) the interspecific hybrids performed better than both the cultivated control and the wild species. In a number of genotypes, glycoalkaloid levels were close to or lower than those of the control varieties, suggesting that selection for low glycoalkaloid content is possible. The results also indicated that glycoalkaloids from S. commersonii may be lost rapidly. Indeed, some hybrids were found to have the same glycoalkaloid profile as S. tuberosum. Finally, the results showed that among the parameters considered, glycoalkaloid content is the most sensitive to variation. Therefore, glycoalkaloid determination should be used for routine control of genotypes produced by interspecific hybridization.     

Genetic diversity and origin of cultivated potatoes based on plastid microsatellite polymorphism

The origin of cultivated potatoes has remaining questions. In this study, 237 accessions of all cultivated species and 155 accessions of wild species closely related to cultivated potatoes, including their putative ancestors, were analyzed using 15 plastid microsatellites (SSRs) to investigate genetic diversity and their relationships with the wild species. We here used polymorphic plastid SSRs we developed from potato plastid genome sequences as well as already known plastid SSR markers. All 15 loci were polymorphic and identified a total of 127 haplotypes. Dramatic decreases in levels of genetic diversity were revealed in landraces in comparison with wild ancestor species. The plastid SSR results showed a decrease in haplotype number and diversity from Peru to both north and south. Phylogenetic analysis revealed two distinct groups. One of them, group A, contained the majority of accessions of cultivated species of the Solanum tuberosum Andigenum group including all accessions of cultivated diploid and triploid cytotypes of this group (S. chaucha, S. phureja, and S. stenotomum by a former taxonomic system) and most of tetraploid accessions of the S. tuberosum Andigenum group (S. tuberosum subsp. andigenum), and the majority of accessions of wild ancestors from the northern members of the S. brevicaule complex. Another group B comprised most of the wild species accessions and almost exclusively hybrid cultivated species which have introgressed plastid genomes from the other wild gene pools. Lack of clustering of traditional cultivated species (as used above) support a revised group classification of S. tuberosum.     

A microsatellite and morphological assessment of the Russian National cultivated potato collection

The germplasm collections of the Vavilov Institute of Plant Industry, Russia represent the first germplasm collection made for potatoes, now numbering 8,680 accessions. It has tremendous historical and practical importance and a rich history, having been used to document a polyploid series in the cultivated species, to formulate initial taxonomic hypotheses in potato, for studies of interspecific hybridization, and serving as the germplasm base for Russian breeding efforts. Despite its importance and size, there has never been a study of its molecular diversity, and there were many gaps in its passport data. The purpose of the present study is to obtain morphological, ploidy, and microsatellite (SSR) data needed to set up a useful subset of the collection of cultivated potatoes and closely related wild species, and to use this collection to study cultivated potato taxonomy and phylogeny. Through assessments of viability, passport data, and chromosome counts, we selected a subset of 238 cultivated and 54 wild accessions. A morphological and nuclear SSR study of these collections distinguished only three cultivated species: Solanum curtilobum, S. juzepczukii and S. tuberosum, not the many more cultivated potato species of prior taxonomic treatments. The SSR study supports the ideas of S. acaule as one of the parental species for S. curtilobum and S. juzepczukii. The morphological and SSR results are very similar to other recent studies of cultivated species, and show the need to reclassify the collection of cultivated potatoes by modern taxonomic criteria.     

A Potato Collecting Expedition in the Province of Jujuy,Argentina and Disease Indexing of Virus and Fungus Pathogens in Andean Cultivars

A potato collecting expedition was carried out in the province of Jujuy, Argentina in March 24 to April 15, 2001. The objective was to collect local cultivars of potatoes and wild potato species, covering high mountain valleys not previously collected or areas where germplasm was not available. A total of 247 accessions was collected, 188 cultivated accessions of S. tuberosum subsp. andigenum, four of Solanum tuberosum subsp. tuberosum, two of S. curtilobum and 53 accessions of wild species. The wild species collected were S. acaule subsp. acaule, S. chacoense, S. infundibuliforme, S. megistacrolobum subsp. megistacrolobum, S. microdontum, S. gourlayi, S. × viirsooi and S. oplocense. Herbarium voucher specimens were obtained when possible. For the collection of cultivated potatoes, tubers were gathered from farmer's fields and in a few cases from stores or markets. Seed samples were generally obtained for the wild species. Detailed collection site data were recorded at every site. After breaking dormancy, the accessions of the cultivated species were screened for the presence of Potato virus Y (PVY), Potato virus X (PVX), Potato leafroll virus (PLRV), Potato virus S (PVS), Potato virus M (PVM), Potato virus V (PVV), Andean potato latent virus (APLV), Andean potato mottle virus (APMoV), Potato rough dwarf virus (PRDV), Potato spindle tuber viroid (PSTVd) and Spongospora subterranea f. sp. subterranea. PSTVd, APLV and PRDV were not detected, but different levels of infection are reported for the other pathogens assayed.     

A recombination-derived mitochondrial genome retained stoichiometrically only among Solanum verrucosum Schltdl. and Mexican polyploid wild potato species

We previously found a specific DNA fragment, designated Band 1, in the cytoplasm of a Mexican hexaploid wild potato species, Solanum demissum, and varieties with the S. demissum cytoplasm. In this study, we show that Band 1 also occurs substoichiometrically in various species and varieties. The S. demissum DNA sequence harboring Band 1 was extended by genome walking to 7,040 bp, and whole-genome sequencing of the S. verrucosum genome generated a 10,794-bp contig with the corresponding sequence. Their 3′-terminal end sequences had 100 % homology with segment 2 of the S. tuberosum mitochondrial genome, proving that Band 1 originated from a recombination-derived mitochondrial genome. Both normal and recombination-derived mitochondrial genomes co-existed in Band 1 carriers. The presence or absence of Band 1 was surveyed for four accessions of tomato and wild relatives and 172 accessions of 38 Solanum species or potato and tuber-bearing wild relatives, mostly from Mexican species. Together, with the results of our previous survey (mostly for South American species), we conclude that Band 1 is retained stoichiometrically only among S. verrucosum and Mexican polyploid species, supporting S. verrucosum as a maternal ancestor for all Mexican polyploid species. The presence or absence of Band 1 was not uniform within these species and was not associated with ploidy, geographical distribution, or latitude. Several evolutionary hypotheses to explain intraspecific variation were discussed.     

2个马铃薯杂交组合F1的SSR鉴定

杂交育种是马铃薯新品种选育的重要方法, 其杂种F₁真实性鉴定是获得目标性状单株的关键环节。为选育优质、高产、抗病性及抗旱性强的马铃薯新品种, 用马铃薯品种"费乌瑞它"(Favorita)分别与"J07-6"和"陇薯3号"杂交, 获得了杂种F1代。本试验利用SSR标记技术对"Favorita"×"J07-6"、"Favorita"×"陇薯3号"2个杂交组合F1共86个单株的真实性进行了鉴定。试验从43对SSR引物中筛选出2对适宜引物STM1049和S7。利用这2对引物进行PCR扩增, 将"Favorita"×"J07-6"杂交种F1和"Favorita"×"陇薯3号"杂交种F1的SSR带型划分为双亲互补型、缺失型、父本型和母本型4种类型。依据SSR带型特征, 从"Favorita"×"J07-6"和"Favorita"×"陇薯3号"2个杂交组合F₁单株中分别鉴定出真杂种34个和27个, SSR分子标记技术用于马铃薯杂交种真实性鉴定是可靠的。该研究结果可为马铃薯杂交种优良株系选育提供依据。     

Patterns of solanidine glycoalkaloid variation in four gene pools of the cultivated potato

Patterns of variation for the solanidine-based glycoalkaloids have been determined for tuber material of genebank accessions of landraces of the cultivated potato and three closely-related wild species. Total levels were low in the cultivated taxa investigated,S. phureja, S. stenotomum, and the tetraploidS. tuberosum in its two formsS. tuberosum ssp.tuberosum andS. tuberosum ssp.andigena. The only solanidine-based glycolkaloids found in the tubers investigated in this study were-solanine and-chaconine, with one additional related compound, dehydrocommersonine, found in tubers of an accession ofS. canasense. The ratio of chaconine : solanine differed markedly from a mean of 0.98 inS. phureja to 2.28 in ChileanS. tuberosum ssp.tuberosum. The chaconine : solanine ratio was relatively constant within these two taxa, reflecting their relatively narrow genetic bases, but was much more variable within the taxa regarded as ancestral,S. stenotomum andS. tuberosum ssp.andigena. The wild speciesS. sparsipilum has been implicated in the origin of the tetraploid potato,S. tuberosum, but there was no support for this from the patterns of variation in chaconine : solanine ratio. The cline of chaconine : solanine ratios in cultivated potatoes along the spine of S. America does not appear to relate to the influence of local wild species through introgression, but may instead reflect differing selective pressures acting in different parts of the native range of the cultivated potato.     

Influence of incorporated wild Solanum genomes on potato properties in terms of starch nanostructure and glycoalkaloid content

Interspecific somatic hybrids produced by protoplast fusion between two wild Solanum species (S. acaule, acl; S. brevidens, brd) and cultivated potato Solanum tuberosum (tbr) were analyzed in terms of the starch nanometer-range structure and glycoalkaloid (GA) contents. The crystallinity of starch granules, the average size of starch crystallites, and the lamellar distances were obtained from tuber samples using wide-angle and small-angle X-ray scattering methods. These measurements showed that incorporation of wild genomes from either nontuberous (brd) or tuberous (acl) Solanum species caused no significant modifications of the nanostructure of potato starch. In contrast, the GA profiles of the hybrids, which were analyzed by LC-ESI-MS in both tuber and foliage samples, differed considerably from those of cultivated potato. Regardless of the low total tuber GA concentrations (approximately 9 mg/100 g of fresh weight), the somatic hybrids contained GAs not detected in the parental species. A high proportion of spirotype GAs consisting of 5,6-dihydrogenated aglycons, for example, alpha-tomatine and tomatidine bound with solatriose, and chacotriose were found in the hybrids. In conclusion, the foliage of interspecific hybrids contained a higher variation in the structures of GAs than did the tubers.     

Adaptive values of wild × cultivated sorghum (<Emphasis Type="Italic">Sorghum bicolor</Emphasis> (L.) Moench) hybrids in generations F<Subscript>1</Subscript>, F<Subscript>2</Subscript>, and F<Subscript>3</Subscript>

Gene flow between cultivated and their wild relatives is one of the main ecological concerns associated with the introduction genetically modified (GM) cultivars. GM sorghum cultivar has been developed and its commercial production may be possible in the near future. The rate of gene flow depends on the fitness of wild × cultivated sorghum hybrids. The study aimed at estimating adaptive values of wild × cultivated sorghum hybrids in generations F₁, F₂, and F₃ compared to their parents. Artificial crosses of four wild sorghums, five cultivated sorghums, and two male sterile lines were made to produce the F₁ generation, which were advanced to F₂ and F₃. Each hybrid generation and their respective parents were evaluated for their adaptive value at two sites in a randomised complete block design with seven replicates. The resulting progenies did not show serious fitness penalties. Some hybrids were as fit as their respective wild parents and no consistent differences exist between the three generations studied. Thus, the resultant wild × cultivated hybrids may act as avenue for introgression.     

Diversity of Nordic Landrace Potatoes (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Revealed by AFLPs and Morphological Characters

The management of genebank collections of cultivated potato is costly due to the need for in vitro maintenance and virus eradication. Therefore, it is important to set up conservation strategies, which prevent duplicates entering the collections. In this study, 32 Nordic potato landraces were studied for 57 morphological traits and analysed for amplified fragment length polymorphism (AFLP). Most landraces could be distinguished based on the morphological characters, except five accessions. Using five primer combinations to generate 114 reproducible AFLPs, of which 63 (55%) were polymorphic, the five morphologically indistinguishable accessions were placed into two groups with identical AFLP patterns, suggesting that some of the accessions were redundant for long-term preservation. The AFLP data showed that the Nordic collection of potato landraces is composed of genetically different clones, and morphological analysis revealed a wide range of variability. This variability seems to be distributed randomly over the Nordic region since the cluster analysis based on AFLPs and morphological traits revealed no grouping based on the country of origin. Principal component analysis suggests that fewer morphological traits than used in this study will be sufficient to discriminate between different genotypes of cultivated potato (Solanum tuberosum L.). Future possibilities for rationalising potato collections are discussed.     

Assessment of artificial selection in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) and Asian rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) using QTL data

Maize (Zea mays L.) and Asian rice (Oryza sativa L.), two most important cereals for human nutrition, have undergone strong artificial selection during a long period of time. Currently, a number of genes with stronger signals of selection have been identified through combining genomic and population genetic approach, but research on artificial selection of maize and Asian rice is scarcely done from the perspective of phenotypic difference of a number of agronomic traits. In this study, such an investigation was carried out on the basis of 179 published studies about phenotypic quantitative trait locus (QTL) mapping of Zea and Oryza species via QTL sign test. At the overall level, the proportions of antagonistic QTLs of Zea and Oryza species were 0.2446 and 0.2382 respectively, deviating significantly from neutrality. It indicated that these two genera have undergone similar selection strength during their evolutionary process. A previous study showed that 4 traits undergoing the directional selection during domestication were identified in Asian rice via QTL sign test, and 16 individual traits in Asian rice and 38 ones in maize that newly detected in this study deviated significantly from neutrality as well, demonstrating the dominant influence of artificial selection on them. Moreover, analysis of different categories of cross type including O. sativa × Oryza rufipogon (perennial and annual forms) crosses, maize × teosinte (Zea mays subsp. parviglumis) crosses, O. sativa × O. sativa crosses, and maize × maize crosses showed that their proportions of antagonistic QTLs were 0.1869, 0.1467, 0.2649, and 0.2618 respectively. These results revealed that selection strength of domestication is significantly stronger than that of modern genetic improvement. However, interestingly, the proportion of antagonistic QTLs (0.1591) in maize × maize with long-term selection was very similar to that (0.1467) in the maize × teosinte (Zea mays subsp. parviglumis) crosses. It suggested that some favorable traits could be cultivated within a few decades if we carry out strong selection. In addition, the proportions of antagonistic QTLs of the widely cultivated hybrids of rice (Minghui 63 × Zhenshan 97) and maize (Zheng 58 × Chang 7-2) in China were 0.309 and 0.3472 respectively. It suggested that selection during modern genetic improvement has significantly acted on them.     

Crossability relationships between the common potato, Solanum tuberosum spp. tuberosum, and its wild diploid relatives S. kurtzianum and S. ruiz-lealii

The common potato, Solanum tuberosum spp. tuberosum (tbr), is a tetraploid species with a narrow genetic base, but with a large number of related species that harbor great genetic diversity for numerous characters of agronomic interest. S. kurtzianum (ktz) and S. ruiz-lealii (rzl) are wild diploid relatives with potential resistance to adverse biotic and abiotic factors. To evaluate if this wild germplasm can be incorporated by conventional crosses into the tbr gene pool, pollen-pistil compatibility relations and seed set in interspecific crosses with tbr were investigated. In 4x tbr × 2x ktz crosses and the reciprocals, 89% and 52.2% of genotypic combinations (respectively) were compatible at the pollen-pistil level. Seeds were obtained from some genotypic combinations in both directions of the cross, suggesting that functional 2n gametes might be produced by particular ktz genotypes. In 2x rzl × 4x tbr crosses and their reciprocals, 35% and 11.7% (respectively) of the genotypic combinations were compatible at the pollen-pistil level, but no seeds were obtained. These results indicate that the reproductive isolation between 4x tbr and 2x ktz is incomplete, and that gene exchange between them is feasible. Moreover, further studies will have to be carried out with rzl to ascertain its crossability with 4x tbr.     

Diversity of the European indigenous wild apple Malus sylvestris (L.) Mill. in the East Ore Mountains (Osterzgebirge), Germany: I. Morphological characterization

Malus sylvestris (L.) Mill. is the only indigenous wild apple species in Europe and recognized as rare and endangered species. An important prerequisite for the preservation of M. sylvestris is the identification of ‘true type’ M. sylvestris individuals because hybrids with cultivated apple could be also occurred. In this study variations among 625 putative M. sylvestris trees originated in the East Ore Mountains were evaluated with 20 selected morphological traits. The importance of these traits concerning the differentiation of ‘true type’ and hybrids of M. sylvestris were determined by statistical analysis. The results demonstrated a strong correlation of individual traits and those contributing mostly to variation of M. sylvestris were detected by the Principal Component Analysis. The relationship of 284 M. sylvestris, 18 M.?×?domestica, M. floribunda 821 and M. robusta 5 individuals were calculated based on the morphological data and presented in a dendrogram. The individuals grouped in two main clusters and 68 hybrids of M. sylvestris were identified. The main result of the morphological data of these hybrids indicates that the grouping is mainly based on the flower and leaf pubescence and less on the fruit size.     

The extent of embryo and endosperm growth following interspecific hybridization between Cicer arietinum L. and related annual wild species

In vivo interspecific pollinations were performed and immature seed development investigated by histological methods in order to study crossability barrier(s) in Cicer L. species wide hybridization. Seven of the eight wild annual Cicer species, belonging to the secondary and tertiary gene pools, were used in reciprocal crosses with the cultivated chickpea. It was confirmed that the zygote was formed in all interspecific crosses. The embryos showed continued and retarded growth at different rate in various crosses, but eventually aborted at an early pro-embryo stage in all crosses except C. arietinum L. ×C. echinospermum Dav. Reciprocal cross differences were observed in early embryo growth rate and could have implications in obtaining hybrids. This study further emphasizes the necessity for developing appropriate and efficient in vitro procedures for rescuing immature globular hybrid pro-embryos, which will make the wild Cicer gene resources amenable to chickpea improvement.     

Intraspecific and interspecific crossability in three Ziziphus species (Rhamnaceae)

Three cultivated species of Ziziphus Mill. [Z. jujuba Mill., Z. mauritiana Lam. and Z. spina-christi (L.) Willd.] comprising a total of seventeen cultivars/genotypes were selected for a mating system study. Phenological stage determination showed an overlap in blooming between the three species during June. Intraspecific cross compatibility was evaluated for Z. jujuba and Z. mauritiana, and interspecific crosses between the three species were carried out during two flowering seasons. Following intraspecific hand cross-pollination, pollen tube formation and growth to the ovule 24 h after pollination (HAP) were observed in all three species. Although very low seed set was obtained following intraspecific Z. jujuba crosses, histological studies showed normal embryo formation and endosperm development, suggesting that double fertilization had occurred and that embryo development arrest occurred later. Following interspecific hand pollination, in vivo pollen grain germination and the formation of pollen tubes and their growth to the ovule 24 HAP were observed in compatible crosses, which also set fruit and seed. Viable embryos were obtained from Z. mauritiana × Z. jujuba, Z. mauritiana × Z. spina-christi and Z. spina-christi × Z. mauritiana crosses. Putative hybrids from these crosses were germinated in vitro, and the resultant plantlets were studied using flow cytometric analysis. Indications of true hybrid origin were obtained based on total 2C-DNA content of plantlets, but they died during the hardening-off process. This study showed that gene flow among these species is possible, thus increasing the potential that interspecific crosses can be used for genetic crop improvement.     

Synchronous evidence from both phenotypic and molecular signatures for the natural occurrence of sympatric hybridization between cultivated soybean (Glycine max) and its wild progenitor (G. soja)

The genus Glycine subgenus Soja includes two species, the important crop, soybean (G. max), and its wild progenitor (G. soja), and an intermediate semi-wild type (G. gracilis). However, the origin of the semi-wild type is in dispute, from independent evolutionary processes or from sympatric hybridization, although many studies support the hypothesis of a hybridization origin. Here, we reveal conclusive evidence for the natural occurrence of sympatric hybridization between wild and cultivated soybeans leading to G. gracilis by capturing natural F₁ seeds based on phenotypic and molecular signatures. The F₁ hybrids occurred through pollen flow between wild and cultivated soybeans, with many heterozygous loci and similar genetic parameters compared to the maternal wild populations; while paternal soybeans showed the lowest genetic diversity. The F-statistics showed genetic differentiation of the hybrids from the paternal soybeans and maternal wild populations; the differentiation of hybrids from wild soybeans was less than that of hybrids from cultivated soybeans. Natural F₁ hybrids generated various seed sizes in the offspring, suggesting that cultivar-genes introgressed into wild soybeans, and the degree of outcrossing in wild soybeans seems to be very important for gene spread, exchange and recombination in the evolution of populations.     

Diversity of the European indigenous wild apple (Malus sylvestris (L.) Mill.) in the East Ore Mountains (Osterzgebirge), Germany: II. Genetic characterization

In the present study genetic diversity and hybridization with cultivars were investigated in a population of the endangered European wild apple species Malus sylvestris (L.) Mill. with the aim to establish a basis for the implementation of conservation activities and to ensure its long-term preservation. A total of 284 putative M. sylvestris trees located in the East Ore Mountains were investigated along with a standard set of reference apple genotypes proposed by the European Cooperative Program for Plant Genetic Resources (ECPGR) and 13 old apple cultivars often cultivated in Saxony. The genetic analysis was performed using 12 microsatellite markers also recommend by the ECPGR. To differentiate ‘true type’ M. sylvestris individuals, hybrids and apple cultivars (Malus × domestica Borkh.) a model-based cluster analysis was performed using STRUCTURE. Two clusters were identified consisting of M. sylvestris and M. × domestica genotypes. About 40 % of the putative M. sylvestris showed an admixture of the species-specific allele frequencies and were defined as hybrids. The genetic diversity of the ‘true type’ M. sylvestris population was still high but slightly lower than in the apple cultivars especially since some SSR loci were fixed on one or few alleles in the M. sylvestris population. The differentiation parameters between ‘true type’ wild apple and cultivars indicated a clear discrimination between the wild and cultivated apple individuals. This fact confirms our expectation of the existence of ‘true type’ M. sylvestris individuals in the East Ore Mountains and argues for the realization of preservation measures in this area.